Plant Genetic Engineering Laboratory, School of Agriculture and Food Sciences, The University of Queensland, St Lucia, Queensland, Australia.
Nat Protoc. 2020 Nov;15(11):3663-3677. doi: 10.1038/s41596-020-0392-7. Epub 2020 Oct 1.
The complexity of current nucleic acid isolation methods limits their use outside of the modern laboratory environment. Here, we describe a fast and affordable method to purify nucleic acids from animal, plant, viral and microbial samples using a cellulose-based dipstick. Nucleic acids can be purified by dipping in-house-made dipsticks into just three solutions: the extract (to bind the nucleic acids), a wash buffer (to remove impurities) and the amplification reaction (to elute the nucleic acids). The speed and simplicity of this method make it ideally suited for molecular applications, both within and outside the laboratory, including limited-resource settings such as remote field sites and teaching institutions. Detailed instructions for how to easily manufacture large numbers of dipsticks in house are provided. Using the instructions, readers can create more than 200 dipsticks in <30 min and perform dipstick-based nucleic acid purifications in 30 s.
目前的核酸分离方法较为复杂，限制了其在现代实验室环境之外的应用。在这里，我们描述了一种快速且经济实惠的方法，可使用基于纤维素的试纸从动物、植物、病毒和微生物样本中纯化核酸。只需将自制的试纸浸入三种溶液中即可纯化核酸：提取液（用于结合核酸）、洗涤缓冲液（用于去除杂质）和扩增反应液（用于洗脱核酸）。这种方法的速度和简单性使其非常适合分子应用，无论是在实验室内部还是外部，包括远程现场和教学机构等资源有限的环境。文中提供了如何在内部轻松大量生产试纸的详细说明。按照说明，读者可以在 <30 分钟内制作出 200 多个试纸，并在 30 秒内完成基于试纸的核酸纯化。
